Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, P...Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 mug/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.展开更多
3-Hydroxypropionaldehyde (3-HPA) is a potential valuable chemical and new broad-spectrum antim-icrobial substance. In order to improve the conversion of 3-HPA/glycerol, our work studied the two-step process from gly...3-Hydroxypropionaldehyde (3-HPA) is a potential valuable chemical and new broad-spectrum antim-icrobial substance. In order to improve the conversion of 3-HPA/glycerol, our work studied the two-step process from glycerol to 3-HPA, and investigated the influence of cell harvest time, glycerol concentration, biomass con-centration, pH and temperature on the production of 3-HPA by Lactobacillus reuteri CG001, respectively. The re-sults showed that molar conversion yield of 3-HPA/glycerol reached 97.9% under the condition that 200 mmol·L-1 glycerol was converted by 25.3 g·L-1 resting cell for 1 h at 30 ℃. The cells could not be reused directly because the L. reuteri almost lost its bioconversion activity completely, but the ability of glycerol conversion could gradually recover if the fresh medium was added to the deactivated cell for 4 h.展开更多
文摘Expression vector p301-bG1 contains a Sw gene and a bialaphos resistance gene both driven by glyceraldehydes-3-phosphate dehydrogenase (GPD) gene promoter isolated from Lentinus edodes ( Berk.) Sing. Using p301-bG1, PEG-mediated transformation of protoplast of L. edodes was studied. Mixed with PEG-purified plasmid DNA, the protoplasts of L. edodes were treated with PEG solution and cultured on CYM regeneration plate containing 40 mug/mL bialaphos. Bialaphos-resistant and GUS-positive transformants were obtained using this transformation system. Although the transformation efficiency was relatively low, the protocols release large expenses on expensive instrument and restriction enzymes, providing a simple and economical method for mushroom breeding at the molecular level.
基金Supported by the National-Natural Science Foundation of China (20906035) and the Fundamental Research Funds for the Central Universities (JB-JX 1002).
文摘3-Hydroxypropionaldehyde (3-HPA) is a potential valuable chemical and new broad-spectrum antim-icrobial substance. In order to improve the conversion of 3-HPA/glycerol, our work studied the two-step process from glycerol to 3-HPA, and investigated the influence of cell harvest time, glycerol concentration, biomass con-centration, pH and temperature on the production of 3-HPA by Lactobacillus reuteri CG001, respectively. The re-sults showed that molar conversion yield of 3-HPA/glycerol reached 97.9% under the condition that 200 mmol·L-1 glycerol was converted by 25.3 g·L-1 resting cell for 1 h at 30 ℃. The cells could not be reused directly because the L. reuteri almost lost its bioconversion activity completely, but the ability of glycerol conversion could gradually recover if the fresh medium was added to the deactivated cell for 4 h.